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Plasma miRNA markers and application thereof

A technology of markers and plasma, applied in the field of plasma miRNA markers, preparation kits and kits, can solve the problem of large trauma

Pending Publication Date: 2021-11-30
SHENZHEN BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] (3) Non-invasiveness of detection
Although microRNA in tumor tissue can be used as a new marker for tumor diagnosis and metastasis prediction, the operation or puncture to collect microRNA in tumor tissue is relatively invasive.

Method used

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  • Plasma miRNA markers and application thereof
  • Plasma miRNA markers and application thereof
  • Plasma miRNA markers and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] Example 1 Screening of miRNAs associated with confirmed HCV infection

[0110] 1.1 Collection of samples and collation of sample data

[0111] The inventors collected anti-HCV positive and HCV RNA positive blood donor samples at Shenzhen Blood Center from January to June 2014 (the samples used for research were collected, sampled, subpackaged, and kept under uniform conditions during the same period). By sorting out the sample data, the inventor selected 5 samples that were positive for HCV RNA and had an anti-HCV COI value greater than or equal to 3.8 for miRNA chip detection.

[0112] Among them, the statistics of anti-HCV positive blood donors in this study are shown in Table 7.

[0113] Table 7:

[0114]

[0115] 1.2 Isolation and purification of plasma miRNA

[0116] 1.2.1 Plasma preparation: 2ml of EDTA-anticoagulated peripheral venous blood was collected from all enrolled patients and healthy controls, and processed within 2 hours at room temperature. Bloo...

Embodiment 2

[0131] Embodiment 2QPCR verifies the miRNA of differential expression

[0132] According to the detection results of the miRNA chip, miRNA-20 and miRNA-122 were selected for large sample QPCR verification.

[0133] 133 anti-HCV positive samples and 56 healthy controls were selected according to the method of sample collection and sample data arrangement in Example 1.

[0134] 2.1 The RNA extraction process is the same as in Example 1.

[0135] 2.2 Reverse transcription of miRNA to synthesize cDNA:

[0136] Each miRNA reverse transcription stem-loop primer is designed according to the specific sequence of the miRNA in miRBase, and the total RNA is reverse transcribed into the corresponding cDNA by a two-step method. The reverse transcription stem-loop primers designed respectively for miRNA-20 and miRNA-122 have the nucleotide sequences shown in SEQID NO:3,5. The inventor simultaneously performed QPCR for quantitative analysis of cel-miR-39 with no expression difference. The...

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Abstract

The invention provides a group of plasma miRNA (micro Ribonucleic Acid) markers, the plasma miRNA markers are used for determining HCV (Hepatitis C Virus) infection, and the markers comprise miR-20 and miR-122. Through differential expression analysis of miRNA in plasma, the inventor finds that the plasma miRNA marker combination is used for confirmation and diagnosis of anti-HCV+ / HCV RNA+ (anti-HCV positive and HCV RNA positive) blood donors, anti-HCV+ / HCV RNA-(anti-HCV positive, HCV RNA negative) blood donors, and anti-HCV- / HCV RNA+ (anti-HCV negative and HCV RNA positive) blood donors, and has good sensitivity, specificity and universality.

Description

technical field [0001] The present invention relates to the field of biological detection, specifically, the present invention relates to plasma miRNA markers and applications thereof, more specifically, the present invention relates to plasma miRNA markers, the use of miR-20 and miR-122 as plasma miRNA markers, and the use of reagents in Use in the preparation of the kit and the kit. Background technique [0002] Hepatitis C virus (HCV) is mainly transmitted through blood, and the high prevalence of HCV in western countries makes it the cause of 90-95% post-transfusion hepatitis. In order to ensure blood safety, blood collection and supply institutions in various countries use anti-HCV detection as a routine screening item for HCV infection in blood donors. According to the latest national standards of the People's Republic of China "GB18467-2011 Blood Donor Health Examination Requirements" and "GB18469-2012 Whole Blood and Component Blood Quality Requirements", there are ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12Q1/6851
CPCC12Q1/6883C12Q1/6851C12Q2600/158C12Q2600/178C12Q2525/207C12Q2563/107C12Q2525/301C12Q2521/107
Inventor 王立林曾劲峰杨宝成朱为刚卢亮洪文旭叶贤林熊文古醒辉郑欣许晓绚陈云龙曾雪珍崔秀娟刘衡邬林枫杜丹丹李彤彭龙
Owner SHENZHEN BLOOD CENT
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